Forgrave Lauren M, Wang Meng, Yang David, DeMarco Mari L
Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, Canada.
Department of Pathology and Laboratory Medicine, St. Paul's Hospital, Providence Health Care, 1081 Burrard St, Vancouver, V6Z 1Y6, Canada.
Pract Lab Med. 2021 Nov 27;28:e00260. doi: 10.1016/j.plabm.2021.e00260. eCollection 2022 Jan.
The term "proteoforms" describes the range of different structures of a protein product of a single gene, including variations in amino acid sequence and post-translational modifications. This diversity in protein structure contributes to the biological complexity observed in living organisms. As the concentration of a particular proteoform may increase or decrease in abnormal physiological states, proteoforms have long been used in medicine as biomarkers of health and disease. Notably, the analytical approaches used to analyze proteoforms have evolved considerably over the years. While ligand binding methods continue to play a large role in proteoform measurement in the clinical laboratory, unanticipated or unknown post-translational modifications and sequence variants can upend even extensively tested and vetted assays that have successfully made it through the medical regulatory process. As an alternate approach, mass spectrometry-with its high molecular selectivity-has become an essential tool in detection, characterization, and quantification of proteoforms in biological fluids and tissues. This review explores the analytical techniques used for proteoform detection and quantification, with an emphasis on mass spectrometry and its various applications in clinical research and patient care including, revealing new biomarker targets, helping improve the design of contemporary ligand binding diagnostics, and as mass spectrometric laboratory developed tests used in routine patient care.
“蛋白质异构体”一词描述了单个基因的蛋白质产物的不同结构范围，包括氨基酸序列的变化和翻译后修饰。蛋白质结构的这种多样性导致了在生物体中观察到的生物复杂性。由于特定蛋白质异构体的浓度在异常生理状态下可能会增加或减少，长期以来蛋白质异构体在医学上一直被用作健康和疾病的生物标志物。值得注意的是，多年来用于分析蛋白质异构体的分析方法有了很大的发展。虽然配体结合方法在临床实验室的蛋白质异构体测量中仍然发挥着重要作用，但意外的或未知的翻译后修饰和序列变异甚至可能颠覆那些经过广泛测试和审查、并已成功通过医学监管程序的检测方法。作为一种替代方法，具有高分子选择性的质谱已成为检测、表征和定量生物体液和组织中蛋白质异构体的重要工具。本综述探讨了用于蛋白质异构体检测和定量的分析技术，重点是质谱及其在临床研究和患者护理中的各种应用，包括揭示新的生物标志物靶点、帮助改进当代配体结合诊断方法的设计，以及作为常规患者护理中使用的质谱实验室开发检测方法。
